Developing species-specific primers to identify Bulinus truncatus and Bulinus beccari, the intermediate hosts of Schistosoma haematobium in Saudi Arabia

• Published in: Gene Vol. 499; no. 2; pp. 256 - 261
• Main Authors: Mostafa, Osama M.S., Bin Dajem, Saad M., Al-Qahtani, Ahmed, Ibrahim, Essam H., Al-Quraishy, Saleh A.S.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.05.2012
• Subjects: Animals; Bulinus - genetics; Bulinus - parasitology; Bulinus beccari; Bulinus truncatus; DNA; DNA Primers; genes; Genetic variability; Host-Parasite Interactions; intermediate hosts; nucleotide sequences; Polymerase Chain Reaction - methods; random amplified polymorphic DNA technique; Random Amplified Polymorphic DNA Technique - methods; RAPD-PCR; Saudi Arabia; Schistosoma haematobium; sequence analysis; snails; Species Specificity; Species-specific primer; Saudi Arabia; B. truncatus; Schistosoma haematobium; Genetic variability; Bulinus truncatus; RAPD; S. haematobium; Species-specific primer; S. japonicum; KSA; RAPD-PCR; Bulinus beccari; PCR; RFLP; S. mansoni; B. beccari
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2012.03.024

This work aimed to determine the inter- and intra-specific variations in populations of Bulinus truncatus and Bulinus beccari, the intermediate hosts of Schistosoma haematobium in Saudi Arabia, and to develop species-specific primers to identify these snails as a first step in the development of multiplex PCR for simultaneously identifying the snails and diagnosing its infections in a single step. Two populations of B. truncatus were collected from Asser and Bisha (A and B), and two B. beccari populations were collected from Mahial Asser and Merba (C and D). The snails’ genomic DNA was extracted and amplified using 5 different primers. The primers displayed variable intra- and inter-specific differences across the populations. The largest RAPD-PCR fragments were cloned into a vector as a preparatory step for sequencing. Similarity searches for the sequenced cloned inserts revealed no similar sequences in the GenBank database or its associated databases. Specific primers used to target the B. truncatus and B. beccari genomes were designed using the Gene Runner program and based on the DNA sequences obtained from RAPD fragment sequence analyses. Using these primers for specific PCRs resulted in expected single-band PCR products of 536bp for B. beccari and 478bp for B. truncatus. These results will be helpful for simultaneously identifying B. truncatus and B. beccari snails and diagnosing S. haematobium infections within the snails using single step multiplex PCR. ► Specific primers designed to target the B. truncatus and B. beccari genomes. ► Using these primers for PCRs resulted in a single-band PCR product for both snails. ► These results will be helpful for identifying B. truncates and B. beccari snails.